Affinage

HOOK3

Protein Hook homolog 3 · UniProt Q86VS8

Length
718 aa
Mass
83.1 kDa
Annotated
2026-06-10
20 papers in source corpus 13 papers cited in narrative 13 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

HOOK3 is a coiled-coil cargo adaptor that bridges organelle membranes to microtubule motors, scaffolding bidirectional transport and Golgi organization (PMID:11238449, PMID:31320392). Its bipartite architecture pairs a conserved N-terminal domain that binds microtubules and the dynein light intermediate chain 1 (LIC1) with a divergent C-terminal region that engages organelle membranes, including the cis-Golgi (PMID:11238449, PMID:27482052). The crystallized Hook domain contacts LIC1 through two conserved surface residues required to assemble a stable dynein-dynactin ternary complex, and a separate region of HOOK3 allosterically activates processive dynein-dynactin motility (PMID:27482052). HOOK3 simultaneously binds the autoinhibitory stalk/tail of the kinesin-3 KIF1C, releasing KIF1C autoinhibition and raising its microtubule landing rate, so that a single HOOK3 scaffold links minus-end (dynein) and plus-end (KIF1C) motors for opposed transport (PMID:31320392, PMID:31217419, PMID:40312563). This dual-motor scaffolding drives defined cargo events: KIF1C acts non-canonically as an adaptor that, via HOOK3 binding the lysosome-anchored RUFY3, activates dynein-driven retrograde lysosomal transport (PMID:39394274). HOOK3 also organizes membrane-associated and centrosomal compartments: its C-terminal region binds the scavenger receptor A cytoplasmic domain to control SR-A turnover (PMID:17237231), and it interacts with PCM1 to localize to pericentriolar satellites and support interkinetic nuclear migration during cortical neurogenesis (PMID:20152126). During mitosis, sequential phosphorylation by ERK1c and AuroraA detaches HOOK3 from microtubules and enhances its GM130 association to drive Golgi fragmentation (PMID:34189435). A HOOK3-RET chromosomal fusion that retains the HOOK3 coiled-coil and the RET kinase domain is oncogenic in transformation and xenograft assays (PMID:17639057).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 2001 High

    Established HOOK3's bipartite domain logic — that one terminus reads microtubules and the other reads organelle membranes — defining it as a candidate physical link between the cytoskeleton and the Golgi.

    Evidence In vitro binding, fractionation, immunofluorescence and overexpression in cell culture

    PMID:11238449

    Open questions at the time
    • Did not identify the motor or membrane receptors mediating each interaction
    • Mechanism of Golgi fragmentation on overexpression unresolved
  2. 2003 Medium

    Showed HOOK3 function can be hijacked by a pathogen, as the Salmonella effector SpiC binds HOOK3 and phenocopies its dominant-negative disruption of Golgi and lysosome distribution.

    Evidence GST pulldown, reciprocal Co-IP, and dominant-negative expression in macrophages

    PMID:12950921

    Open questions at the time
    • Single lab
    • Did not map the HOOK3 region bound by SpiC
    • Link to specific trafficking machinery not defined
  3. 2004 Medium

    Connected HOOK3 to interferon-induced immune effectors by showing the GTPase IIGP binds it in a nucleotide-dependent manner at Golgi membranes.

    Evidence Yeast two-hybrid and Co-IP from IFNγ-stimulated macrophages with fractionation

    PMID:15075236

    Open questions at the time
    • Functional consequence of the interaction not established
    • Single lab, two methods
  4. 2007 High

    Defined a membrane-receptor cargo for HOOK3 at the residue level, showing its basic C-terminal region binds acidic residues of the SR-A cytoplasmic tail to regulate receptor stability and turnover.

    Evidence Yeast two-hybrid, GST pulldown, truncation mutants, and siRNA functional readouts

    PMID:17237231

    Open questions at the time
    • Whether SR-A turnover depends on motor-driven transport not tested
    • Endosomal/lysosomal routing of SR-A unresolved
  5. 2007 Medium

    Demonstrated a disease-relevant gain of function, as a HOOK3-RET chromosomal fusion fuses HOOK3 coiled-coils to the intact RET kinase and is oncogenic.

    Evidence 5'RACE, Western blot, NIH3T3 transformation and nude mouse xenograft

    PMID:17639057

    Open questions at the time
    • Mechanism of RET activation by HOOK3 coiled-coils (e.g. dimerization) not dissected
    • Single lab
  6. 2010 High

    Placed HOOK3 at pericentriolar satellites via PCM1 and tied this to a developmental process, linking the interaction to interkinetic nuclear migration and neural progenitor maintenance.

    Evidence Co-IP plus in vivo dominant-negative disruption and live imaging of cortical neurogenesis

    PMID:20152126

    Open questions at the time
    • Motor requirement for satellite trafficking not defined here
    • Direct vs indirect PCM1 binding not resolved
  7. 2015 Low

    Implicated HOOK3 in endosomal transport kinetics with a downstream consequence for amyloid precursor protein processing.

    Evidence siRNA knockdown, live-cell endosomal transport assay, β-amyloid ELISA

    PMID:25799409

    Open questions at the time
    • Single functional assay per endpoint with limited mechanistic detail
    • Cargo and motor mediating the endosomal effect unidentified
  8. 2016 High

    Resolved how HOOK3 engages dynein, showing the crystallized Hook domain binds LIC1 through two conserved residues required for stable dynein-dynactin complex formation and identifying a separate region needed for motility activation.

    Evidence Crystal structure, structure-based mutagenesis, in vitro binding and single-molecule motility assays

    PMID:27482052

    Open questions at the time
    • How membrane cargo binding couples to activation not shown
    • Regulation of the activating region unresolved
  9. 2019 High

    Recast HOOK3 as a dual-motor scaffold, reconstituting a ternary complex in which it activates dynein-dynactin and simultaneously binds the autoinhibitory KIF1C tail to enable opposed transport.

    Evidence In vitro reconstitution with purified proteins, single-molecule motility, mass spectrometry, and KIF1C recruitment in cells; converging with landing-rate assays showing release of KIF1C autoinhibition

    PMID:31217419 PMID:31320392

    Open questions at the time
    • What determines net directionality on a given cargo not defined
    • In vivo cargoes of the dual-motor complex not enumerated here
  10. 2021 Medium

    Identified the mitotic switch controlling HOOK3, with sequential ERK1c and AuroraA phosphorylation detaching it from microtubules and boosting GM130 binding to fragment the Golgi.

    Evidence Kinase substrate assay, phospho-site mutagenesis, Co-IP, microtubule co-sedimentation, Golgi imaging

    PMID:34189435

    Open questions at the time
    • Phospho-sites and their effect on motor binding not fully mapped
    • Single lab
  11. 2024 Medium

    Revealed a non-canonical adaptor role for KIF1C, where it activates dynein-driven retrograde lysosomal transport through HOOK3 and RUFY3 without using its own motor activity.

    Evidence Co-IP, siRNA knockdown, live-cell lysosome imaging, motor-dead and dominant-negative KIF1C

    PMID:39394274

    Open questions at the time
    • How directionality is biased toward dynein in this context not fully resolved
    • Single lab
  12. 2025 High

    Defined the HOOK3-KIF1C interface at atomic resolution and showed it is necessary and sufficient for anterograde transport, while placing PTPN21 at the same KIF1C tail as a regulator.

    Evidence Crystal structure of Hook3(553-624)–KIF1C(714-809), structure-based mutagenesis, Co-IP, and live-cell transport assays in RPE1 cells

    PMID:40312563

    Open questions at the time
    • How PTPN21 and HOOK3 binding to the shared site are coordinated not resolved
    • Structural basis for simultaneous dynein and KIF1C engagement not shown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How HOOK3 selects, integrates, and directionally biases its competing dynein and KIF1C motors on a given membrane cargo in vivo remains unresolved.
  • No structure of HOOK3 simultaneously bound to dynein-dynactin and KIF1C
  • Regulatory logic linking phosphorylation, RUFY3/GM130/SR-A cargo identity, and motor choice unintegrated

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 4 GO:0008092 cytoskeletal protein binding 2 GO:0098772 molecular function regulator activity 2
Localization
GO:0005794 Golgi apparatus 2 GO:0005764 lysosome 1 GO:0005768 endosome 1 GO:0005815 microtubule organizing center 1 GO:0005829 cytosol 1
Pathway
R-HSA-5653656 Vesicle-mediated transport 3 R-HSA-9609507 Protein localization 3 R-HSA-1266738 Developmental Biology 1
Partners
DYNC1LI1GM130KIF1CMSR1PCM1PTPN21RETRUFY3
Complex memberships
dynein-dynactin-Hook3 activated transport complexpericentriolar satellite

Evidence

Reading pass · 13 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2001 Hook3 contains a conserved NH2-terminal domain that mediates microtubule binding and a divergent COOH-terminal domain that mediates organelle (Golgi membrane) binding. Hook3 bound to Golgi membranes in vitro and was enriched at the cis-Golgi in vivo. Overexpression of Hook3 caused fragmentation of the Golgi complex. In vitro binding assays, subcellular fractionation, overexpression/dominant-negative in cell culture, immunofluorescence The Journal of cell biology High 11238449
2003 Salmonella SpiC protein directly binds Hook3 (demonstrated by GST-SpiC pulldown and co-immunoprecipitation from infected macrophages), and SpiC expression phenocopies a Hook3 dominant-negative mutant by disrupting Golgi morphology and altering lysosome distribution, indicating SpiC targets Hook3 function to alter cellular trafficking. GST pulldown, co-immunoprecipitation, dominant-negative expression in Vero cells and macrophages Molecular microbiology Medium 12950921
2004 The IFN-inducible 47 kDa GTPase IIGP physically interacts with Hook3 in a GTP-bound conformation-dependent manner, as shown by yeast two-hybrid and co-immunoprecipitation from IFNγ-stimulated macrophages; both proteins co-localize in Golgi-membrane-enriched fractions. Yeast two-hybrid, co-immunoprecipitation, subcellular fractionation Journal of cell science Medium 15075236
2007 Hook3 interacts directly with the cytoplasmic domain of scavenger receptor A (SR-A); the positively charged C-terminal Val614-Ala717 region of Hook3 binds the negatively charged residues Glu12, Asp13, and Asp15 of the SR-A cytoplasmic domain. Hook3 knockdown (siRNA) increased total and surface expression, ligand uptake, and protein stability of SR-A without affecting synthesis or maturation, indicating Hook3 participates in SR-A turnover. Yeast two-hybrid, mass spectrometry, GST pulldown, co-immunoprecipitation, co-sedimentation, siRNA knockdown, truncation mutants The Journal of biological chemistry High 17237231
2007 The HOOK3-RET fusion gene, resulting from chromosomal rearrangement fusing HOOK3 exon 11 to RET exon 12, produces an 88 kDa chimeric protein retaining HOOK3 coiled-coil domains and the intact RET tyrosine kinase domain. Expression of HOOK3-RET cDNA in NIH3T3 cells caused transformed foci formation and tumor formation in nude mice, confirming oncogenic activity. 5'RACE, Western blot, NIH3T3 transformation assay, nude mouse xenograft Endocrine-related cancer Medium 17639057
2010 Hook3 interacts with PCM1 (Pericentriolar Material 1) to recruit Hook3 to pericentriolar satellites, enabling trafficking of pericentriolar satellite components. Disruption of the Hook3-PCM1 interaction in vivo impairs interkinetic nuclear migration in embryonic neural progenitors, leading to overproduction of neurons and premature depletion of the neural progenitor pool in the developing neocortex. Co-immunoprecipitation, in vivo dominant-negative disruption, live imaging of interkinetic nuclear migration, cortical neurogenesis assays Neuron High 20152126
2016 The conserved Hook domain of Hook3 directly interacts with the dynein light intermediate chain 1 (LIC1). Crystal structure of the Hook domain was solved, and structure-based mutagenesis identified two conserved surface residues critical for LIC1 binding; Hook proteins with mutations in these residues fail to form a stable dynein-dynactin ternary complex. A separate region of Hook3 is specifically required for allosteric activation of processive dynein-dynactin motility. Crystal structure determination, structure-based mutagenesis, in vitro binding assays, single-molecule motility assays The Journal of cell biology High 27482052
2019 Hook3 acts as a scaffold for both cytoplasmic dynein-1/dynactin and kinesin-3 KIF1C, forming a ternary complex in vitro with purified components. Full-length Hook3 binds to and activates dynein/dynactin motility, and also binds to the KIF1C tail region without activating KIF1C motility. This scaffolding allows dynein to transport KIF1C toward the microtubule minus end and KIF1C to transport dynein toward the plus end. In cells, KIF1C can recruit Hook3 to the cell periphery. In vitro reconstitution with purified proteins, single-molecule motility assays, mass spectrometry, cell biology (KIF1C recruitment assay) The Journal of cell biology High 31320392
2019 Hook3 binds to the stalk/tail region of KIF1C (the same region that mediates KIF1C autoinhibition) and increases the landing rate of KIF1C onto microtubules in vitro, functioning as a cargo adaptor that releases KIF1C autoinhibition to enable cargo-activated transport. In vitro microtubule binding/landing assays with purified proteins, co-immunoprecipitation, KIF1C truncation analysis Nature communications High 31217419
2021 ERK1c phosphorylates HOOK3 early in mitosis, and a subsequent phosphorylation by AuroraA is also required. These phosphorylations cause HOOK3 to detach from microtubules and increase its interaction with GM130, leading to Golgi destabilization and fragmentation during mitosis. Substrate identification (kinase assay), phosphorylation-site mutagenesis, co-immunoprecipitation, microtubule co-sedimentation assay, cell imaging of Golgi fragmentation iScience Medium 34189435
2024 KIF1C supports retrograde lysosomal transport (toward the microtubule minus end) driven by dynein through interaction with Hook3, which associates with the lysosome-anchored protein RUFY3. KIF1C motor activity is not required and in fact inhibits this process; instead, KIF1C functions non-canonically as an adaptor to activate dynein-driven lysosomal transport via Hook3. Co-immunoprecipitation, siRNA knockdown, live-cell imaging of lysosome transport, dominant-negative and motor-dead KIF1C constructs Communications biology Medium 39394274
2025 Crystal structure of the Hook3(553-624)–KIF1C(714-809) complex was determined, and structure-based mutagenesis showed that this complex formation is necessary and sufficient for full-length protein interaction in HEK293T cells and for Hook3- and KIF1C-mediated anterograde transport in RPE1 cells. PTPN21 also interacts with the same KIF1C tail region to regulate transport. Crystal structure determination, structure-based mutagenesis, co-immunoprecipitation in HEK293T cells, live-cell cargo transport assays in RPE1 cells EMBO reports High 40312563
2015 Hook3 deficiency in cultured cells slows endosomal transport and increases β-amyloid production, establishing a functional role for Hook3 in endosomal trafficking that modulates amyloid precursor protein processing. siRNA knockdown, live-cell endosomal transport assay, β-amyloid ELISA PloS one Low 25799409

Source papers

Stage 0 corpus · 20 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2001 The Golgi-associated hook3 protein is a member of a novel family of microtubule-binding proteins. The Journal of cell biology 167 11238449
2016 Assembly and activation of dynein-dynactin by the cargo adaptor protein Hook3. The Journal of cell biology 103 27482052
2010 Hook3 interacts with PCM1 to regulate pericentriolar material assembly and the timing of neurogenesis. Neuron 98 20152126
2019 Hook3 is a scaffold for the opposite-polarity microtubule-based motors cytoplasmic dynein-1 and KIF1C. The Journal of cell biology 66 31320392
2019 PTPN21 and Hook3 relieve KIF1C autoinhibition and activate intracellular transport. Nature communications 57 31217419
2007 HOOK3-RET: a novel type of RET/PTC rearrangement in papillary thyroid carcinoma. Endocrine-related cancer 55 17639057
2003 The Salmonella SpiC protein targets the mammalian Hook3 protein function to alter cellular trafficking. Molecular microbiology 52 12950921
2004 IIGP, a member of the IFN inducible and microbial defense mediating 47 kDa GTPase family, interacts with the microtubule binding protein hook3. Journal of cell science 28 15075236
2015 Hook proteins: association with Alzheimer pathology and regulatory role of hook3 in amyloid beta generation. PloS one 21 25799409
2019 miR-496 remedies hypoxia reoxygenation-induced H9c2 cardiomyocyte apoptosis via Hook3-targeted PI3k/Akt/mTOR signaling pathway activation. Journal of cellular biochemistry 19 31436348
2007 The microtubule-binding protein Hook3 interacts with a cytoplasmic domain of scavenger receptor A. The Journal of biological chemistry 16 17237231
2023 The Circ_35953 induced by the NF-κB mediated the septic AKI via targeting miR-7219-5p/HOOK3 and IGFBP7 axis. Journal of cellular and molecular medicine 13 36974922
2022 Identification of a novel HOOK3-FGFR1 fusion gene involved in activation of the NF-kappaB pathway. Cancer cell international 11 35081975
2021 Mitotic HOOK3 phosphorylation by ERK1c drives microtubule-dependent Golgi destabilization and fragmentation. iScience 9 34189435
2021 Midazolam increases cisplatin-sensitivity in non-small cell lung cancer (NSCLC) via the miR-194-5p/HOOK3 axis. Cancer cell international 9 34321010
2024 KIF1C facilitates retrograde transport of lysosomes through Hook3 and dynein. Communications biology 8 39394274
2024 HOOK3 suppresses proliferation and metastasis in gastric cancer via the SP1/VEGFA axis. Cell death discovery 6 38228617
2026 Durable response to selpercatinib in HOOK3-RET fusion positive, α-fetoprotein producing metastatic pancreatic ductal adenocarcinoma with intestinal-type differentiation. Oncology letters 0 42238005
2025 Molecular basis for assembly and activation of the Hook3 - KIF1C complex-dependent transport machinery. EMBO reports 0 40312563
2025 Exosomal miR-3529-3p increases the sensitivity of trastuzumab via Wnt/β-catenin signaling pathway by targeting HOOK3. International immunopharmacology 0 40614603

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